Mitochondrial p32-mediated regulation of p-TBK1 affects the radiosensitivity of colorectal cancer.

IF 5.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Life sciences Pub Date : 2025-07-12 DOI:10.1016/j.lfs.2025.123854

Yanjin Wu, Haitao Zhou, Jiahui Meng, Wenyue Zhao, Xiaotong Zhao, Yujia Hou, Qin Wang, Feng Wang, Qiang Liu, Yang Liu

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引用次数: 0

Abstract

Purpose: Many colorectal cancer (CRC) patients respond poorly to radiotherapy due to radioresistance. Understanding the molecular mechanisms underlying this resistance is crucial. It was demonstrated that p32, a mitochondrial protein translation regulator, is related to cancer development. However, its specific function and mechanism in CRC, has not yet been investigated. This study aims to explore the role of p32 in CRC and its impact on radiotherapy sensitivity.

Methods: Cell viability was evaluated by MTT and EdU assay. Mitochondrial DNA (mtDNA) leakage was quantified by RT-qPCR. Radiosensitivity was indicated by cellular phosphorylation of H2AX (γH2AX) foci, phosphorylation of ataxia telangiectasia mutated (p-ATM) and phosphorylation of checkpoint kinase 2 (p-CHK2) levels, as well as by mice tumor model subjected to radiotherapy. Moreover, histological and transcriptomic analysis of p32 expression were performed in CRC patients.

Results: In p32-KO cells, we observed reduced cell viability, damaged mitochondria, mtDNA leakage, and increased radiosensitivity. Furthermore, depletion of p32 induced the DNA damage response (DDR) by activating cytoplasmic DNA sensing cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING)-TANK-binding kinase 1 (TBK1), which was reversed by p32/TBK1 double knockout. Depletion of p32 also induced the mitochondrial fragmentation, induced mtDNA leakage through the mitochondrial permeability transition pore (mPTP), effects that could be mitigated by Mdivi-1 or Cyclosporin A (CsA).

Conclusions: Our study demonstrates that inhibiting p32 in CRC enhances radiosensitivity by causing mitochondrial dysfunction, increasing mitochondrial fission, inducing mtDNA leakage and activating the cGAS-STING-TBK1 pathway. These findings provide a potential therapeutic target for overcoming radioresistance in CRC.

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线粒体p32介导的p-TBK1调控影响结直肠癌的放射敏感性。

目的：许多结直肠癌（CRC）患者由于放射耐药而对放疗反应较差。了解这种耐药性背后的分子机制至关重要。研究表明，线粒体蛋白翻译调节因子p32与癌症的发生有关。然而，其在结直肠癌中的具体功能和机制尚不清楚。本研究旨在探讨p32在结直肠癌中的作用及其对放疗敏感性的影响。方法：采用MTT法和EdU法测定细胞活力。RT-qPCR法测定线粒体DNA （mtDNA）渗漏量。通过细胞中H2AX （γH2AX）病灶磷酸化、共济失调毛细血管扩张突变（p-ATM）磷酸化和检查点激酶2 （p-CHK2）磷酸化水平以及放疗小鼠肿瘤模型显示放射敏感性。此外，在结直肠癌患者中进行了p32表达的组织学和转录组学分析。结果：在p32-KO细胞中，我们观察到细胞活力降低，线粒体受损，mtDNA泄漏，放射敏感性增加。此外，p32的缺失通过激活细胞质DNA感应环GMP-AMP合成酶(cGAS)-干扰素基因刺激因子(STING)- tank结合激酶1 （TBK1）诱导DNA损伤反应（DDR），该反应通过p32/TBK1双敲除而逆转。p32的缺失还会导致线粒体断裂，诱导mtDNA通过线粒体通透性过渡孔（mPTP）渗漏，Mdivi-1或Cyclosporin A （CsA）可以减轻这种影响。结论：我们的研究表明，在CRC中抑制p32可通过引起线粒体功能障碍、增加线粒体裂变、诱导mtDNA泄漏和激活cGAS-STING-TBK1通路来增强放射敏感性。这些发现为克服结直肠癌的放射耐药提供了一个潜在的治疗靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Life sciences 医学-药学

CiteScore

12.20

自引率

1.60%

发文量

841

审稿时长

6 months

期刊介绍： Life Sciences is an international journal publishing articles that emphasize the molecular, cellular, and functional basis of therapy. The journal emphasizes the understanding of mechanism that is relevant to all aspects of human disease and translation to patients. All articles are rigorously reviewed. The Journal favors publication of full-length papers where modern scientific technologies are used to explain molecular, cellular and physiological mechanisms. Articles that merely report observations are rarely accepted. Recommendations from the Declaration of Helsinki or NIH guidelines for care and use of laboratory animals must be adhered to. Articles should be written at a level accessible to readers who are non-specialists in the topic of the article themselves, but who are interested in the research. The Journal welcomes reviews on topics of wide interest to investigators in the life sciences. We particularly encourage submission of brief, focused reviews containing high-quality artwork and require the use of mechanistic summary diagrams.